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Molecular Biotechnology Programme Uppsala University School of Engineering UPTEC X 02 013 Date of issue 2002-04
Author
Torun Engfeldt
Title (English)
Optimisation of mRNA in situ technique using non-radioactive probes
Title (Swedish): Optimering av mRNA in situ teknik med icke radioaktiva prober
Abstract
The sensitivity of in situ hybridisation using non-radioactive labelling was investigated and improved using digoxigenin-labelled RNA probes. In situ hybridisation with digoxigenin labelled RNA probes gives rise to a significant, specific signal and very low background.
Subjecting paraffin-embedded tissue sections to microwave heating prior to hybridisation significantly increased the signal intensity. Time requirements for detection of hybridised dig- labelled probe was minimised to 3 days, compared to 2-3 weeks for radioactive labelling. The method proved to be reliable and applicable when studying the expression of Fractalkine, Fractalkine receptor, Target N and N receptor in rat spinal cord and brain.
Keywords
In situ hybridisation, digoxigenin, non- radioactive probes, gene expression Supervisor
Katarina Nordqvist
AstraZeneca R&D Södertälje, Sweden
Examiner
Bengt Fundin
AstraZeneca R&D Södertälje, Sweden
Project name Sponsors
Language
English
Security
ISSN 1401-2138 Classification
Supplementary bibliographical information Pages
34
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